Figure 6. Specific expression of netrin-1 after doxycycline treatment in stable HeLa cell lines.

Stable HeLa cell lines (Flp-In TRex tetracycline transactivator HeLa cells transfected with the 3 netrin-1 constructs harboring the NTN1 mutations or the WT construct) were grown for 24 hours in the presence or absence of doxycycline. The experiments were replicated 3 times. Confocal microscopy (original magnification, ×63) showed that no netrin-1 was detected in the absence of doxycycline. None of the proteins was detected in the nucleus. Hoechst staining (blue) and anti–c-Myc (green, to detect netrin-1) were used. In each condition, arrowheads point to the same cell at low (first line) and high (second line) magnification. Scale bars: 60 μm for the top row and 9.5 μm for the bottom 2 rows.